Formulation for soft anticholinergic analogs

ABSTRACT

Topical formulations comprising soft glycopyrrolates are useful for treating excessive sweating conditions in subjects, such as humans suffering from hyperhidrosis. Preferably, at least one soft anticholinergic agent is provided in an effective amount or concentration in an anhydrous formulation that can inhibit excessive perspiration resulting from a condition such as hyperhidrosis.

BACKGROUND

Various anticholinergic compounds and formulations for those compoundshave been previously described. Muscarinic receptor antagonists arefrequently used therapeutic agents that inhibit the effects ofacetylcholine by blocking its binding to muscarinic cholinergicreceptors at neuroeffector sites on smooth muscle, cardiac muscle, andgland cells as well as in peripheral ganglia and in the central nervoussystem (CNS). However, their side effects, which can include dry mouth,photophobia, blurred vision, urinary hesitancy and retention,drowsiness, dizziness, restlessness, irritability, disorientation,hallucinations, tachycardia and cardiac arrhythmias, nausea,constipation, and severe allergic reactions, often limit their clinicaluse. Topical administration of anticholinergic agents to targeted areas,such as sweat glands, where the localized blockage of muscarinicreceptors will be of clinical benefit, would be a desirable therapeuticstrategy. However, currently used topical anticholinergics can exhibitunwanted systemic side effects which can limit the dosage that can besafely administered.

Glycopyrrolate is among the quaternary ammonium anticholinergics whichhave reduced CNS-related side effects as they cannot cross theblood-brain barrier; however, because glycopyrrolate is eliminatedmainly as unchanged drug or active metabolite, its topicaladministration is often associated with common undesirableanticholinergic systemic side effects. To increase the therapeutic indexof anticholinergics, the soft drug approach has been applied in a numberof different designs starting from various lead compounds, but there isa need for yet other new soft anticholinergics with clinicallymeaningful biological activity. These novel muscarinic antagonists, justas all other soft drugs, are designed to elicit their intendedpharmacological effect at the site of application, but to be quicklymetabolized into their designed-in, inactive metabolite upon enteringthe systemic circulation and rapidly eliminated from the body, resultingin reduced systemic side effects and an increased therapeutic index.

Soft anticholinergic zwitterions have been described in US PatentPublication No. 2012/0141401 (now U.S. Pat. No. 8,568,699), and itsrelated patents, U.S. Pat. Nos. 8,071,693; 7,538,219; and 7,417,147.Soft anticholinergic esters have been described in US Patent PublicationNo. 2012/0177590 (now U.S. Pat. No. 8,628,759) and its related patentsU.S. Pat. Nos. 8,147,809; 7,576,210; and 7,399,861. Although thesepublished applications and patents identified the potential for thezwitterion or ester forms of anticholinergics to be used for treatinghyperhidrosis, the fact that activity and duration of action againsthyperhidrosis are unexpectedly high herein, based on a comparison topublished mydriasis data, was not known or previously described.

Each of the US Patent Publication Nos. 2012/0141401 (U.S. Pat. No.8,568,699) and 2012/0177590 (U.S. Pat. No. 8,628,759), and their relatedpatents U.S. Pat. Nos. 8,147,809; 8,071,693; 7,576,210; 7,538,219;7,417,147; and 7,399,861 are hereby incorporated by reference in theirentireties.

Hyperhidrosis is an idiopathic pathological condition characterized byexcessive, uncontrollable sweating beyond that required to cool thebody. A hyperfunction of the sweat glands and a disturbance of theircholinergic stimulation have been described as possible causes of thiscondition. It is known to affect approximately 3% of the population.Hyperhidrosis not only may result in intense social embarrassment, butalso may even interfere with a person's occupation.

Hyperhidrosis most often involves one or several areas, especially thehands, axillae, feet or face, although it can even involve the wholebody. Axillary hyperhidrosis is the most common form, followed by palmarhyperhidrosis. Antiperspirants alone are generally not effective intreating this excessive perspiration. Oral medications are occasionallybeneficial, but may have side effects. Other therapeutic alternativesinclude surgical procedure such as endoscopic thoracic sympathectomy.Although the surgery affords permanent benefit in some 40% to 90% ofaffected individuals, it is invasive, requires general anesthesia and isnot without potential side effects. As many as 50% of persons who haveundergone thoracic sympathectomy develop compensatory and annoyingsweating of the trunk or thighs. Botulinum A neurotoxin (BOTOX), whichblocks the action on sweat glands of acetylcholine that is released bythe autonomic nerves, has proven effective in hyperhidrosis. Minuteamounts of BOTOX injected into the palms or axillae of affectedindividuals result in statistically significant benefit. The effectlasts for several months but requires repeated injections and is oftennot a suitable alternative for pediatric patients. Iontophoresis haslimited efficacy and cannot be used for axillary areas.

A non-invasive, convenient and effective treatment having high sweatreduction activity, long duration and with fewer side effects would be awelcome alternative for treating hyperhidrosis. An improved method oftreating hyperhidrosis has recently been described in copending U.S.patent application Ser. No. 14/213,242, filed Mar. 14, 2014 (inventorsBODOR and ANGULO), incorporated by reference herein in its entirety.

Topical formulations comprising soft anticholinergic analogs, such assoft ester analogs of glycopyrrolate, have been previously proposed foruse in treating hyperhidrosis; however, stable, pharmaceuticallyacceptable formulations of such esters which can meet regulatoryrequirements or provide commercially viable shelf-life for such productshave been elusive. Thus, what is needed in the art is a stable,pharmaceutically acceptable, and commercially viable formulation for atopically administered composition comprising a soft anticholinergicanalog.

SUMMARY

The subject application concerns topical formulations for treatingexcessive sweating conditions in subjects, such as humans suffering fromhyperhidrosis. A composition herein comprises at least one softanticholinergic agent, which is a soft ester analog of glycopyrrolate,in an effective amount or concentration that can inhibit excessiveperspiration resulting from a condition such as hyperhidrosis. Oneembodiment is a topical composition comprising: (a) at least onecompound having the formula (1):

said compound having the R, S, or RS stereoisomeric configuration at the2 position and 1′ and 3′ positions, or being a mixture thereof, and (b)anhydrous ethanol, provided that said topical composition is anhydrous.

One preferred embodiment of a topical composition comprises: (a) atleast one compound having the following stereospecific formula (2):

said compound having the R stereoisomeric configuration at the 2position and having the R, S, or RS stereoisomeric configuration at the1′ and 3′ positions (designated by asterisks), or being a mixturethereof, and (b) anhydrous ethanol, provided that said topicalcomposition is anhydrous.

Another embodiment provides a topical pharmaceutical compositioncomprising (a) one or more compounds of the foregoing formula (2), (b)anhydrous ethanol and (c) one or more pharmaceutically acceptablecarriers or excipients, provided that said topical composition isanhydrous. Yet another embodiment provides a topical compositioncomprising (a) and (b) above; (c) optionally, at least one gelling orviscosity controlling ingredient; and (d) optionally at least oneadditional carrier or excipient; provided that said topical compositionis anhydrous and comprises from about 1% to about 25% of the compound offormula (2), said composition having greater storage stability comparedto a composition comprising an aqueous solvent or aqueous buffer.

Methods of treating or inhibiting or ameliorating excessive sweating,including conditions such as hyperhidrosis, using a topical compositionas described herein, are also included. The methods of copending U.S.patent application Ser. No. 14/213, 242, filed Mar. 14, 2014, are ofparticular interest and advantage when carried out by administering atopical formulation comprising an ethyl ester of formula (2) above andanhydrous ethanol, provided that said topical formulation is anhydrous.

A composition of the subject application can be formulated as a solid orsemi-solid, powder, gel, cream, lotion, foam, solution, suspension,aerosol, patch, wipes or emulsion, or the like, and is formulated fortopical application for the treatment, inhibition or amelioration ofhyperhidrosis. More preferably, a composition as defined above isformulated as an anhydrous ethanol topical gel, which can providecertain advantages, including superior stability or increased shelf-lifefor the composition, as well as the benefit of minimizing or eliminatingthe need for a separate preservative in the composition.

Additional advantages for a topical anhydrous ethanol gel compositionherein include properties such as fast drying time, limited residue onthe skin or clothing, and facilitation of a capability to be dispensedin metered amounts of product per application. A particular formulationcan further mask stickiness properties that some soft anticholinergics,such as certain compounds described herein, may have.

One formulation comprises about 0.1% to about 30% of the compound in70-99.9% of the non-aqueous solvent, ethanol. The formulation canfurther include one or more additional carriers or excipients, includinga gelling or viscosity controlling excipient, which itself is anhydrous,that is non-aqueous.

The compounds of formulas (1) and (2) are ethyl esters. As esters, thesecompounds are subject to transesterification, which is the process ofexchanging the alkyl group of the ester with the alkyl group of analcohol/alkanol. This reaction is catalyzed by acid or base or evenenzymatically. Unfortunately, transesterification can lead to aninterchange of a significant amount of the drug's ester group for a lessdesirable, less biologically acceptable group. For example, use ofanhydrous methanol as solvent for the ethyl ester leads to unacceptableformation of a significant amount of methyl ester mixed with ethylester. Use of anhydrous ethanol, on the other hand, leads only toformation of ethyl ester as a product of transesterification. Further,by using anhydrous ethanol, and by making certain that the compositionitself is anhydrous, it is possible to avoid hydrolysis of the activeingredient's ethyl ester group.

There is thus provided in one aspect herein a method for treating,inhibiting or ameliorating hyperhidrosis in a subject which comprises:

-   (A) providing a topical composition comprising: (a) from about 1% to    about 25% of a compound having the formula (1):

said compound having the R, S or RS stereoisomeric configuration at the2 position and 1′ and 3′ positions, or being a mixture thereof; (b)anhydrous ethanol; (c) optionally, at least one gelling orviscosity-controlling ingredient; and (d) optionally, at least oneadditional carrier or excipient; provided that said topical compositionis anhydrous; and

-   (B) topically administering the composition to a subject suffering    from excessive sweating, such as hyperhidrosis.

There is further provided in another aspect herein a method fortreating, inhibiting or ameliorating hyperhidrosis in a subject whichcomprises:

-   (A) providing a topical composition comprising: (a) from about 1% to    about 25% of a compound having the formula (2):

said compound having the R stereoisomeric configuration at the 2position and the R, S, or RS stereoisomeric configuration at the 1′ and3′ positions, or being a mixture thereof, (b) anhydrous ethanol; (c)optionally, at least one gelling or viscosity-controlling ingredient;and (d) optionally, at least one additional carrier or excipient;provided that said topical composition is anhydrous; and

-   (B) topically administering the composition to a subject suffering    from excessive sweating, such as hyperhidrosis.

Advantageously, the method can provide reduction of excessive sweatingfor up to about 48 hours. Moreover, surprisingly, topical administrationof the composition can unexpectedly provide a reduction in sweatproduction, as compared to baseline conditions, for at least about six(6) hours by an amount which is substantially equivalent to thereduction of sweat production resulting from administration of acomposition comprising an equivalent concentration of glycopyrrolate,also compared to baseline conditions. Soft ester analogs ofglycopyrrolate were previously believed to require up to 5-10 times theconcentration of glycopyrrolate to provide substantially equivalentactivity.

A preferred method of treating hyperhidrosis in a subject in need ofsame or for treating, inhibiting or ameliorating excessive sweatingtherein, comprises administering the instant composition in accord withthe methods of U.S. patent application Ser. No. 14/213,242. In accordtherewith, the composition as defined herein comprising a compound offormula (2) above is administered to skin of a subject suffering fromhyperhidrosis, before bedtime, such that, compared to untreated,baseline conditions, sweat production is reduced by at least 25% for atleast six (6) hours; and such that sweat production is reduced by anamount substantially equivalent to an amount that sweat production isreduced as compared to untreated, baseline conditions, followingadministration of a composition comprising the same concentration ofglycopyrrolate, and with an improved safety profile compared to topicalglycopyrrolate. In particular, at 5% drug concentration, no systemicanticholinergic side effects were observed for the soft ester in testingdescribed in the '242 application. Also, no systemic anticholinergicside effects were observed in clinical studies at 5% or 10% as describedhereinbelow.

The present method is preferably carried out by topically administeringthe composition to a human subject, to the skin of the subject at asuperficial anatomic area in need of sweat reduction. Preferably, theanatomic area for application or administration of the composition isselected from a hand palm area, a foot plantar area, a groin area, anaxilla area, and a facial area of the subject.

The subject method can reduce sweat production by about 25% to about99%, preferably by about 30% to about 90%, more preferably by at least50%, which can be a clinically significant endpoint for an indicationfor treating hyperhidrosis.

As previously described, the method can employ the compositionformulated as a solid or semisolid, powder, gel, cream, lotion, foam,solution, suspension, aerosol, patch, wipes or emulsion, or the like andcan comprise from about 0.1% to about 30% concentration of the compound,preferably from about 1% to about 25% concentration of the compound,more preferably about 1% to about 20% concentration of the compound, andmost preferably about 2% to about 10% concentration of the compound offormula (1) above, preferably of formula (2).

A method in accordance with the present description can comprisetopically administering to a subject as needed (prn), a composition asdefined herein. Administrations are preferably at least one time perweek, more preferably at least three to four times per week (e.g., everyother day), or can be administered more frequently such as once daily(QD), for example, before bedtime (typically, at night) or after thesubject awakens (typically in the morning, and preferably after a bathor shower); twice daily (BID), e.g., every 10-12 hours; thrice daily(TID), e.g., every 6-9 hours; four times daily (QID), e.g., every 3-5hours; with a preferred upper limit of about 6-8 doses or applicationsper day.

Surprisingly, the subject method, after single or multiple applications,can reduce sweat production for a period of from about 4 hours to about24 hours, and preferably for a period of from about 6 hours to about 12hours.

A preferred composition herein comprises:

-   one or more soft glycopyrrolate analogs of formula (1) or (2) as    active ingredient; and-   anhydrous ethanol (as non-aqueous solvent for the active    ingredient);-   provided that said composition is anhydrous.

As described herein, the subject formulation is preferably a gel.Accordingly a more preferred composition comprises:

-   one or more soft glycopyrrolate analogs of formula (1) or (2) as    active ingredient; anhydrous ethanol (as non-aqueous,    pharmaceutically acceptable solvent for the active ingredient); and-   one or more gelling or viscosity-controlling agents,-   provided that said formulation is anhydrous.

The soft glycopyrrolate analog of formula (1) or (2) is a softanticholinergic ethyl ester. The use of the matching non-aqueous solventethanol avoids mixtures of esters which can result fromtransesterification when an alcohol such as methanol is used as solventfor the ethyl ester. Moreover, the absence of water results in muchgreater storage stability.

Advantageously, anhydrous ethanol can provide for a self-preservingcomposition, which can provide microbial stability to the compositionwithout added preservatives.

Anhydrous ethanol can also inhibit bacterial growth and providedeodorant properties to the composition.

A further advantage of a composition according to the presentdescription is provided by the fact that the non-aqueous solvent,anhydrous ethanol, is volatile, especially at localized temperaturesgenerated by body heat so that, when it is topically applied to asubject, it provides a rapidly drying composition.

A preferred gelling or viscosity controlling agent can be a modifiedcellulose, e.g., hydroxypropyl cellulose (HPC), such as the commerciallyavailable KLUCEL™, which can preferably provide viscosity of thecomposition of from about 100 to about 10,000 cps.

DETAILED DESCRIPTION

Throughout this specification, the following definitions, generalstatements and illustrations are applicable.

The patents, published applications and scientific literature referredto herein establish the knowledge of those with skill in the art and arehereby incorporated by reference in their entireties to the same extentas if each was specifically and individually indicated to beincorporated by reference. Any conflict between any reference citedherein and the specific teachings of this specification shall beresolved in favor of the latter. Likewise, any conflict between anart-understood definition of a word or phrase and a definition of theword or phrase as specifically taught in this specification shall beresolved in favor of the latter.

As used herein, whether in a transitional phrase or in the body of aclaim, the terms “comprise(s)” and “comprising” are to be interpreted ashaving an open-ended meaning. That is, the terms are to be interpretedsynonymously with the phrases “having at least” or “including at least”.When used in the context of a process, the term “comprising” means thatthe process includes at least the recited steps, but may includeadditional steps. When used in the context of a composition, the term“comprising” means that the composition includes at least the recitedfeatures or components, but may also include additional features orcomponents.

The terms “consists essentially of” or “consisting essentially of” havea partially closed meaning, that is, they do not permit inclusion ofsteps or features or components which would substantially change theessential characteristics of a process or composition; for example,steps or features or components which would significantly interfere withthe desired properties of the compounds or compositions describedherein, i.e., the process or composition is limited to the specifiedsteps or materials and those which do not materially affect the basicand novel characteristics of the process or composition.

The terms “consists of” and “consists” are closed terminology and allowonly for the inclusion of the recited steps or features or components.

As used herein, the singular forms “a,” “an” and “the” specifically alsoencompass the plural forms of the terms to which they refer, unless thecontent clearly dictates otherwise.

The term “about” is used herein to mean approximately, in the region of,roughly, or around. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth. In general, the term“about” or “approximately” is used herein to modify a numerical valueabove and below the stated value by a variance of 20%.

As used herein, the recitation of a numerical range for a variable isintended to convey that the variable can be equal to any values withinthat range. Thus, for a variable which is inherently discrete, thevariable can be equal to any integer value of the numerical range,including the end-points of the range. Similarly, for a variable whichis inherently continuous, the variable can be equal to any real value ofthe numerical range, including the end-points of the range. As anexample, a variable which is described as having values between 0 and 2,can be 0, 1 or 2 for variables which are inherently discrete, and can be0.0, 0.1, 0.01, 0.001, or any other real value for variables which areinherently continuous.

In the specification and claims, the singular forms include pluralreferents unless the context clearly dictates otherwise. As used herein,unless specifically indicated otherwise, the word “or” is used in the“inclusive” sense of “and/or” and not the “exclusive” sense of“either/or.”

Technical and scientific terms used herein have the meaning commonlyunderstood by one of skill in the art to which the present descriptionpertains, unless otherwise defined. Reference is made herein to variousmethodologies and materials known to those of skill in the art. Standardreference works setting forth the general principles of pharmacologyinclude Goodman and Gilman's The Pharmacological Basis of Therapeutics,10th Ed., McGraw Hill Companies Inc., New York (2001).

As used herein, “treating” means reducing, hindering or inhibiting thedevelopment of, controlling, inhibiting, alleviating and/or reversingthe symptoms in the individual to which a composition comprising acompound of formula (1) or (2) has been administered, as compared to thesymptoms of an individual not being administered the compound orcomposition. A practitioner will appreciate that the combinations,compositions, dosage forms and methods described herein are to be usedin concomitance with continuous clinical evaluations by a skilledpractitioner (physician or veterinarian) to determine subsequenttherapy. Such evaluation will aid and inform in evaluating whether toincrease, reduce or continue a particular treatment dose, and/or toalter the mode of administration.

The subject compounds or compositions can also prevent the symptoms, orprevent the occurrence of the symptoms, in the individual to which acomposition comprising a compound of formula (1) or (2) above has beenadministered, as compared to the symptoms of an individual not beingadministered the compound or composition. This is not a prevention ofhyperhidrosis or excessive sweating in the absolute sense, it does notprevent the medical condition, as it does not even address thecondition's cause; rather it inhibits the manifestation of the conditionfor the period of time (hours) for which the administered dose iseffective.

The methods described herein are intended for use with anysubject/patient that may experience their benefits. Thus, the terms“subjects” as well as “patients,” “individuals” and “warm-bloodedanimals” include humans as well as non-human subjects, such as animalsthat may experience excessive sweating.

Compounds useful in a composition herein include those of the formula(1):

The compound has the R, S, or RS stereoisomeric configuration at the 2position and at the 1′ and 3′ positions, or is a mixture thereof.

Compounds having the R configuration with respect to chiral center 2 areof particular interest for use in the instant compositions. For example,a preferred compound useful in a composition herein has thestereospecific formula (2):

said compound having the R stereoisomeric configuration at the 2position and the R, S, or RS stereoisomeric configuration at the 1′ and3′ positions (designated by asterisks), or being a mixture thereof.

The following compounds are of particular interest for use in acomposition of the present description:

-   (i)    3-(2-cyclopentylphenylhydroxyacetoxy)-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide;-   (ii)    3-[2(R)-cyclopentylphenylhydroxyacetoxy]-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide;-   (iii)    3′(R)-[2(R)-cyclopentylphenylhydroxyacetoxy]-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide;-   (iv) 3′(S)-[2(R)-cyclopentyl phenyl    hydroxyacetoxy]-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide;-   (v)    1′(R)-3′(S)-[2(R)-cyclopentylphenylhydroxyacetoxy]-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide;-   (vi)1′(S)-3′(S)-[2(R)-cyclopentylphenylhydroxyacetoxy]-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide;-   (vii)    1′(R)-3′(R)-[2(R)-cyclopentylphenylhydroxyacetoxy]-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide; and-   (viii)    1′(S)-3′(R)-[2(R)-cyclopentylphenylhydroxyacetoxy]-1′-methyl-1′-ethoxycarbonylmethylpyrrolidinium    bromide.

It is noted that the above compounds are identical to those originallydisclosed with a correct, but different, naming scheme, in U.S.Provisional Patent Application Ser. No. 61/952,505 filed Mar. 13, 2014.The compounds were previously and respectively disclosed as:

-   (i)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide;-   (ii) (2R)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide;-   (iii) (2R, 3′R)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide;-   (iv) (2R,3′S)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide;-   (v) (2R,1′R,3′S)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide;-   (vi) (2R,1′S,3′S)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide;-   (vii) (2R,1′R,3′R)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide; and-   (viii) (2R,1′S,3′R)    3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidinium    bromide.

The above compounds (i)-(viii) can be used alone or two or more of theabove compounds can be used in combination in a single composition.Various methods of making the instant compounds are described in theart.

An anticholinergically effective amount of such an agent inhibits theeffect of acetylcholine by blocking its binding to muscariniccholinergic receptors at neuroeffector sites. Subjects in need of amethod of eliciting an anticholinergic response are those suffering fromconditions which respond to treatment with an anticholinergic agent,including subjects suffering from excessive sweating or hyperhidrosis.

The compound of formula (1) or (2) is typically administered in the formof a pharmaceutical composition comprising an anticholinergicallyeffective amount of the compound, anhydrous ethanol and a non-toxicpharmaceutically acceptable carrier or excipient, provided that thecomposition itself is also anhydrous. Pharmaceutically acceptablecarriers, or diluents, are well-known in the art. The carriers may beany inert material, organic or inorganic, powders, liquid, or gasessuitable for administration, such as: alcohol such as hexylene glycol,gelatin, gum arabic, lactose, microcrystalline cellulose, starch, sodiumstarch glycolate, calcium hydrogen phosphate, magnesium stearate,talcum, colloidal silicon dioxide, and the like, provided that theingredients are anhydrous.

It has been discovered that the instant formulations, havingadvantageous properties, result when no water or aqueous carrier isadded to the formulation. Thus, a composition herein is an anhydrousformulation. By the term “anhydrous”, is meant the ordinary scientificmeaning of the word, that is, that no water or aqueous excipient isadded to the formulation.

Such compositions may also contain conventional additives such assolvents, stabilizers, wetting agents, emulsifiers, buffers, binders,disintegrants, fragrances, lubricants, glidants, antiadherents,propellants, and the like, just so long as the additives andcompositions are anhydrous, that is, free of water to the extentrequired to avoid significant negative impact on the storage stabilityof the composition (by hydrolysis of the ester drug).

The active ingredient is dissolved in anhydrous ethanol as a solvent, inwhich the compound is soluble or at least slightly soluble. It ispreferred that the apparent pH of the composition be acidic (i.e.apparent pH <7).

The composition herein can be formulated as a solid, semi-solid, orliquid, such as in the form of powders, solutions, lotions, creams,gels, semi-solid sticks, foams, sprays, aerosols, solutions, suspensionsor emulsions, patches, wipes and the like, and is formulated for topicaladministration. By way of illustration only, for treating hyperhidrosis,a topical preparation formulated as an anhydrous antiperspirant stick,gel, spray, cream, solution, foam, emulsion or the like can bepreferred.

In preparing a formulation, it may be necessary to mill the activecompound to provide the appropriate particle size prior to combiningwith the other ingredients. The active compound can be milled to aparticle size of less than 200 mesh.

Some examples of suitable topical carriers or excipients, to be added tothe compound of formula (1) or (2) in absolute ethanol, include alcoholssuch as hexylene glycol and propylene glycol, dimethicone, e.g.dimethicone 350 cSt, dimethicone copolyol, Dimethiconol Blend 20,dimethiconol 20 cSt, cyclomethicone, e.g. cyclomethicone 5-NF, PGE,allantoin, glycerin, vitamin A and E oils, mineral oil, PPG2, myristylpropionate, isopropyl myristate, C₁₂₋₁₅ alkyl lactate, lactose,dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calciumphosphate, alginates, tragacanth, gelatin, calcium silicate,microcrystalline cellulose, polyvinylpyrrolidone, cellulose, and methylcellulose, and mixtures thereof. The formulations can additionallyinclude: lubricating agents such as talc, magnesium stearate, andmineral oil; wetting agents; emulsifying and suspending agents; andpreserving agents such as methyl- and propylhydroxy-benzoates. Thecompositions can be formulated so as to provide quick, modified,sustained or delayed release or activity of the active ingredient afteradministration and/or application to the subject by employing proceduresknown in the art. The use of a separate preserving agent can be avoidedby judicious selection of other ingredients, as discussed in more detailbelow.

The composition may additionally contain one or more optional additivessuch as colorants, perfumes, or the like. In practice, each of theseoptional additives should be compatible with the active compound.Compatible additives are those that do not prevent the use of or resultin the degradation of the compound in the manner described herein.

For purposes of illustration, liquid formulation dosages are expressedbased on a percent solution (g/100 ml) or percent concentration (w/v)unless otherwise stated. For solid formulation dosages, the percentconcentration can be expressed as mg/mg, or w/w concentrations unlessotherwise stated. A person of ordinary skill in the art would readilyunderstand the percent concentration in the context of the type offormulation described.

In general, a therapeutically effective or anticholinergically effectiveamount of a compound of formula (1) or (2) herein is from an about 1%solution (10 mg/ml) to an about 30% solution (300 mg/ml). Preferably,the topical composition dose is from about 1% concentration to about 25%concentration, or more preferably from about 1% concentration to about20% concentration, especially from 2% to 10%, and is most preferredusing a dose application volume of approximately 0.5 to about 1.0 ml or2.0 ml of a composition comprising about 3% to about 6%, e.g., about 5%,of the compound per treated area. The exact dosage of a compound in theinstant composition can vary depending on its potency, the mode ofadministration, the application area, the age and weight of the subjectand the severity of the condition to be treated. The daily dosage may beadministered singly or multiply one to four times daily or more.

Administration prior to bedtime (in accord with a preferred method oftreating hyperhidrosis herein) does not imply at night or a particularhour or time of day; rather, before or prior to bedtime means that thecomposition is preferably administered, generally within about 1-2 hoursprior to a person's normal rest or sleep (typically 4 to 10-hour)period. A before bedtime administration time can provide a preferredresponse or activity of the active compounds of formulas (1) and (2), inaccord with the method of prior copending U.S. patent application Ser.No. 14/213,242.

Administration of a composition as described herein can provide asubstantially identical or similar clinical (sweat reduction) responsein a subject, as compared to administration of a composition containingthe same concentration of glycopyrrolate. Thus, the results of thisdiscovery are surprising in view of previously published mydriaticstudies which suggested that the subject compounds in a composition wererequired to be present in concentration from 5 times to 10 times theconcentration of a glycopyrrolate composition exhibiting a similar orsubstantially identical clinical response.

In addition, administration of a second dose within about 6-10 hoursfollowing the initial dose can also be a preferred method ofadministration or dosing regimen.

The topical composition for treating hyperhidrosis can be a liquidsolution, semi-solid, or solid. Solutions are prepared in the usual way,e.g. with the addition of excipients as well as the anhydrous ethanolsolvent and can include preservatives such as p-hydroxybenzoates, orstabilizers such as alkali metal salts of ethylenediamine tetraaceticacid, optionally using emulsifiers and/or dispersants, and other organicsolvents may optionally be used as solvating agents or dissolving aids,and transferred into vials, ampules, bottles, tubes, syringes, or thelike.

However, the anhydrous composition can have the advantage of minimizing,or eliminating, the need for an additional preservative to be includedin the formulation. Thus, one preferred embodiment of a composition is asubstantially “preservative-free” composition. By “preservative-free” ismeant that the composition, although containing ethanol and evenpossibly another organic solvent which may provide some preservingproperties, has no additional preservative component, added specificallyfor its preservative property to the composition.

Additional carriers or excipients may be used in a composition herein,including, for example, pharmaceutically acceptable organic solventssuch as paraffins (e.g. petroleum fractions), vegetable oils (e.g.groundnut or sesame oil), mono- or polyfunctional alcohols (e.g.hexylene glycol or glycerol), carriers such as natural mineral powders(e.g. kaolins, clays, talc, chalk), synthetic mineral powders (e.g.highly dispersed silicic acid and silicates), sugars (e.g. cane sugar,lactose and glucose), emulsifiers (e.g. lignin, spent sulfite liquors,methylcellulose, starch and polyvinylpyrrolidone) and lubricants (e.g.magnesium stearate, talc, stearic acid and sodium lauryl sulphate).

Compositions herein can be formulated using known techniques, and aregenerally accepted as being formulated with commonly known excipients,including preservatives if needed. For example, the patent literaturedescribes that soft glycopyrrolate compounds are at least partiallywater-soluble. Accordingly, soft glycopyrrolates compounds, such as softanticholinergic analogs (e.g., esters) were earlier described as capableof being formulated in buffer (aqueous or water-based) solutions.However, aqueous components added to the formulation increase thehydrolysis products found in the composition, and decrease the stabilityof the active compound, and consequently decrease the shelf-life of theproduct compared to anhydrous formulations comprising a softanticholinergic analog (ester) as an active ingredient.

Moreover, decreased stability and increased hydrolysis products foundfor a soft anticholinergic analog (ester) formulated in an aqueous orwater-based composition would suggest or even require an addedpreservative to be included in the composition.

In addition to the general preference or need to decrease exposure topreservative chemicals by the subject being treated, certainingredients, such as the antioxidant/pH adjuster ascorbic acid, can haveadditional disadvantages when topically applied. For example, an aqueouspreparation comprising ascorbic acid was found to produce a pink-coloredresidue on the skin of individuals after a few to several hoursfollowing exposure to the preparation.

A preservative-free composition, which is also an ascorbic acid-freecomposition, can therefore provide a further advantage of maintaining acolorless preparation following application and during residence on theskin of a subject. A composition comprising citric acid as anantioxidant/pH adjuster did not result in a pink colored residuefollowing application of the composition to the skin; therefore acomposition herein can include citric acid as an antioxidant.

Experimental data demonstrate that aqueous or water-based compositionsresult in the presence of increased hydrolysis products in thecomposition, and decreased stability of the composition, which leads toreduced shelf-life for a product comprising the composition. Adequateshelf-life is an advantageous factor for regulatory approval, as well ascommercial success for a topical gel composition.

The HPLC experimental data presented in EXAMPLE 1 below also demonstratethe reduction of hydrolysis products Identified, and increased stabilityfor a product comprising an anhydrous topical gel in accordance with thesubject description.

EXAMPLE 1 Proof of Concept

Aqueous, or water-based, topical formulations are the most common inview of the availability of gel-forming components which interact withwater to form hydrogels. The following experiments were conducted usingthe compound, (2R,3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide, (compound (iii) in the above list), which is designated as“BBI-4000” for convenience of reference.

Various formulations of approximately 2% BBI-4000 were made and theirstabilities assessed. The solvent systems were as follows:

-   (a) solvent content: 100% water;-   (b) solvent content: 60% water/40% ethanol;-   (c) solvent content: 30% water/70% ethanol;-   (d) solvent content: 100% ethanol.-   Each sample was assessed at baseline; after 7 days at 25° 0160%    humidity; and after 7 days at 40° C/75%humidity. The percentage    change from baseline for 40° C. after 7 days was calculated in each    case. HPLC analysis was conducted as described in EXAMPLE 2 below.

Testing conclusively showed that, of the four solvent systems tested,only 100% ethanol (i.e. absolute or anhydrous ethanol) was capable ofproviding a composition which essentially maintained the baseline amountof BBI-4000 even after 7 days at the elevated temperature of 40° C.There is clearly a dramatic difference in the stability of the anhydrousethanol formulation compared to the water-containing formulations. Theresults are shown in TABLE I below.

TABLE I Formulation Solvent Content: 100% Water Change from Condition/Day 7 @ Day 7 @ Baseline Timepoint Baseline 25° C./60% 40° C./75% (40°C./75%) BBI-4000 Assay 1.99% 1.91% 1.80% reduction 9.5% BBI-4000 Main 01.90% 7.42% Hydrolysis Products (RRT- 0.79-0.84) Formulation SolventContent: 60% water, 40% ethanol Change from Condition/ Day 7 @ Day 7 @Baseline Timepoint Baseline 25° C./60% 40° C./75% (40° C./75%) BBI-4000Assay 1.99% 1.94% 1.89% reduction 5% BBI-4000 Main 0 0.83% 3.40%Hydrolysis Products (RRT- 0.79-0.84) Formulation Solvent Content: 30%water, 70% ethanol Change from Condition/ Day 7 @ Day 7 @ BaselineTimepoint Baseline 25° C./60% 40° C./75% (40° C./75%) BBI-4000 Assay1.99% 1.95% 1.89% reduction 5% BBI-4000 Main 0 0.84% 3.50% HydrolysisProducts (RRT- 0.79-0.84) Formulation Solvent Content: 100% EthanolChange from Condition/ Day 7 @ Day 7 @ Baseline Timepoint Baseline 25°C./60% 40° C./75% (40° C./75%) BBI-4000 Assay 2.02% 2% 2.01% reduction<1% BBI-4000 Main 0 0 0 Hydrolysis Products (RRT- 0.79-0.84)

EXAMPLE 2 Aqueous Formulations

The following Table II shows the components included in an aqueousformulation comprising BBI-4000, a soft anticholinergic ethyl ester,prepared and subjected to hydrolysis and stability testing:

TABLE II Lot Number (% w/w) BB- BB- BB- BB- BB- Material 61-1 62-1 63-164-1 65-1 BBI-4000 2.00 2.00 2.00 2.00 2.00 Hydroxyethyl Cellulose 1.001.00 1.00 1.00 1.00 Hexylene Glycol 5.00 5.00 5.00 5.00 5.00 BenzylAlcohol 1.00 1.00 1.00 1.00 1.00 Ethanol 95% 26.31 26.32 26.32 26.3226.32 Polysorbate 80 1.00 1.00 1.00 1.00 1.00 Dimethiconol Blend 20 2.502.50 2.50 2.50 2.50 Dibasic Sodium 0.09 0.09 0.09 Phosphate, DriedMonobasic Sodium 0.53 0.53 0.53 Phosphate, Anhydrous Citric Acid,Anhydrous 0.20 Trisodium Citrate 1.16 Dihydrate Water 61.19 60.56 60.5660.56 59.83 2N HCl to pH to pH to pH to pH to pH 5 4.5 5 5.5 5 2N NaOHto pH to pH to pH to pH to pH 5 4.5 5 5.5 5

An HPLC method was developed at a commercial laboratory for assaying thesoft anticholinergic analog, and related substances (includinghydrolysis products):

Apparatus

-   High performance liquid chromatography (HPLC) system Chromatography    data system-   XBridge Shield RP18,4.6×150 mm, 3.5 μm HPLC column-   Analytical balance capable of weighing to 0.00001 g-   Ultrasonic bath-   Volumetric flasks, 1, 5 mL-   Syringe Filter: 25 mm, 0.45 μm, HPF Millex-HV, Millipore or suitable    alternative

Reagents, Supplies, Media and Solutions:

-   BBI-4000 standard-   Water, HPLC grade-   Acetonitrile (can), Optima grade-   Trifluroacetic acid (TFA), Fisher-   Mobile Phase “A”: 0.1% TFA in Water-   Mobile Phase “B”: 0.1% TFA in Acetonitrile-   Auto Sampler Flush: 1:1 Water:Acetonitrile-   Diluent: Acetonitrile    BBI-4000 Standard Preparation (2 mg/mL in Diluent):

The standards were prepared in duplicate by weighing 2.0±0.1 mg ofBBI-4000 into 1 mL volumetric flasks. Dissolved and diluted to volumewith acetonitrile and mixed by inversion.

Sample Preparation (BBI-4000 Gels):

Gel samples were prepared in duplicate at a target concentration of 2mg/mL in a 5-mL volumetric flask. Added 1.5 ml H₂O and mixed to dispersethe sample. Diluted to volume with acetonitrile and filtered an aliquotthrough a syringe filter.

HPLC Conditions:

The liquid chromatographic system was set-up as follows:

-   HPLC Column: XBridge Shield RP18, 4.6×150 mm, 3.5 μm-   Column Temp.: 25±1° C.-   Sample Temp.: ambiant-   Flow Rate: 1.5 mL/min-   Injection Volume: 10 μL-   UV Detection: 220 nm-   Run Time: 20 minutes

The HPLC assay was conducted on formulations at differing pH values, andthe results were reviewed for “Time-Zero” and at 7 days at 40° C.:

The BBI-4000 content was determined. By Day 7, the assay numberdecreased, indicating hydrolysis of the BBI-4000 and some hydrolyticdegradation products were noticeably increased (the two zwitterionstereoisomers, identified by RRT 0.84 and RRT 0.80), indicating lack ofstability of this formulation system. Adjustment of pH, by itself,although providing a lower percent of hydrolytic degradation in thebuffered formulation, did not resolve the issue.

A second experiment was conducted using a preparation comprising 2% of asoft glycopyrrolate ethyl ester (SGE) in an aqueous buffer system, whichwas tested for stability at refrigerated, 25° C. (RT), and 40° C., for 7days, and showed the same trend or similar results.

Thus, independent of pH, when water or aqueous buffer is present, theSGE is relatively rapidly degraded by hydrolysis and is substantiallyreduced in less than one week.

EXAMPLE 3 Anhydrous Formulations

For preparing an anhydrous formulation, it is noted that no water oraqueous solution is added to the preparation.

The anhydrous formulations are based on: ethanol (solvent), hexyleneglycol (moisturizer), and hydroxypropyl cellulose (HPC, gelling agent),in varying amounts or ratios. Each formulation was given anidentification number as follows:

-   69-1=without antioxidant-   73-2=without antioxidant but with polysorbate 80-   72-2=adding propylene glycol and polysorbate 80-   78-1 and 78-2=different quantities of HPC-   79-1=with ascorbic acid as antioxidant/acidifying agent-   79-2=with Vitamin E as antioxidant-   84-1=with citric acid as antioxidant/acidifying agent    Repeat-dose Studies up to 14 days

A 14-day dermal and systemic toxicity and toxicokinetics study inGöttingen Minipigs was conducted and completed using a formulation basedon Formulations 79-1 and 84-1, above, but having a relatively highconcentration of the active drug for testing tolerability. Specifically,the composition of the preparation used in this study included BBI-4000as active ingredient (except in the vehicle-only control), hydroxypropylcellulose as a gelling agent, hexylene glycol as an emollient, ascorbicacid or citric acid as antioxidant/pH adjuster and ethanol as theanhydrous vehicle.

Three groups of one male and one female animal were included in the mainstudy, Group 1 receiving vehicle, Group 2 receiving BBI-4000 gel at 10%concentration and Group 3 receiving BBI-4000 gel at 20% concentration.All groups received 2 mL of gel formulation, once a day, for 14consecutive days, applied to approximately 10% of their body surfacearea on their backs.

The study included daily observations of the site of application andscoring of erythema and edema (if present), daily general examinationsincluding heart rate as well as pupil size assessment at days 1, 2, 3,5, 7, 10 and 14. The frequent observations of heart rate and pupil sizewere intended to identify any potential systemic anticholinergic effect.Main organs were evaluated during necropsy and histopathology evaluationwas completed for treated and untreated skin. Blood samples forchemistry and hematology analysis were collected as well as PK samples.

The results indicated that the composition was well-tolerated, there wasno evidence of erythema or edema in the treated skin of any of theanimals. Daily observation did not report any abnormality in heart rateor any other parameter. Pupil size assessments were reported as normalat all times in all animals. Blood chemistry and hematology parameterswere reported within normal ranges. The necropsy did not reveal anyabnormalities in any of the animals.

Histopathology analysis for the skin treated with an anhydrouscomposition comprising BBI-4000 was unremarkable and identical tonon-treated and vehicle treated skin. All skin samples from thedifferent groups were similar, with minor nonspecific changes that donot appear to be related to treatment. Mild, superficial inflammationreported in the dermis of most skin samples from all groups and from thenon-treated areas suggests this finding is not drug or compositionrelated, but associated with the caging of the animals.

The estimated BBI-4000 dose applied to the skin in this study was 40mg/kg/day for Group 3 and 20 mg/kg/day for Group 2.

The PK analysis revealed variable, dose related systemic exposure ofBBI-4000. The highest concentration was observed at 2 hours after Day-14dosing in a minipig receiving the 20% BBI-4000 concentration. Most ofthe PK values for the carboxylic acid metabolite were below the lowestlimit of quantification (LLOQ=4.75 ng/mL for this assay), consistentwith the short half-life of this metabolite. Group 1 (vehicle) did notreport any value above the LLOQ, as expected.

It was noted during the study that a reddish formulation residue wasobserved in the skin of all animals receiving the ascorbicacid-containing formulation. Although the residue could be removed withwiping from the skin, this type of residue would not be acceptable to ahuman subject; therefore, additional formulations were evaluated. A newexperiment was conducted in 2 new pigs with a new formulation removingthe ascorbic acid, adding citric acid instead. Testing of the citricacid-containing formulation was also well tolerated and no reddish orpink-colored residue was observed.

The following formulations, shown in Table III, were tested forstability:

TABLE III A 84-1 B 84-2 C 84-3 % % % Component (w/w) (w/w) (w/w)BBI-4000 10 10 10 KLUCEL ™ MF 1.25 1.25 1.25 (Hydroxypropyl Cellulose)Hexylene Glycol 10 10 10 DIMETHICONOL 2.5 2.5 2.5 BLEND 20 BHT — 0.1 —Propyl Gallate — — 0.05 Citric Acid, Anhydrous 0.1 0.1 0.1 Ethanol (200proof) 76.15 76.05 76.1 (Anhydrous Ethanol)

KLUCEL™ MF hydroxylpropyl cellulose (HPC) is available commercially froma variety of sources. DOW CORNING® DIMETHICONOL BLEND 20 is a uniqueblend of silicone gum (6%) in dimethicone. BHT is butylatedhydroxytoluene also known as dibutylhydroxytoluene.

BBI-4000 and non-BBI-4000 levels determined at time “zero” are shown inTable IV, below:

TABLE IV Day 0 Results BB-84-1 BB-84-2 BB-84-3 BBI- Assay  9.81%  9.89% 9.72% 4000 (Wt %) TAN % 98.19% 95.15% 92.17% RRT Area % RRT Area % RRTArea % Non- RRT 0.67% BBI- 0.80 4000 by RRT 0.10% RRT 0.62% RRT 6.07%HPLC 0.96 0.80 0.64 (%) RRT 0.86% RRT 0.07% RRT 0.69% 1.09 0.96 0.80 RRT0.19% RRT 0.79% RRT 0.09% 1.48 1.09 0.96 RRT 0.16% RRT 0.81% 1.49 1.09RRT 0.90% RRT 0.17% 2.05 1.49 RRT 2.31% 2.07 Total  1.82%  4.85%  7.83%Non- BBI- 4000 by HPLC

BBI-4000 and non-BBI-4000 levels determined at 7 days, under acceleratedconditions, 40° C., are shown in Table V, below:

TABLE V Day 7 Results BB-84-1 BB-84-2 BB-84-3 BBI- Assay 4000 (Wt 10.32%10.18% 10.08% %) TAN 97.89% 94.75% 93.84% % RRT Area % RRT Area % RRTArea % Non- RRT 0.59% RRT 0.42% RRT 4.28% BBI- 0.80 0.80 0.64 4000 byRRT 0.03% RRT 0.16% HPLC 0.82 0.91 (%) RRT 0.17% RRT 0.15% RRT 0.58%0.91 0.96 0.80 RRT 0.29% RRT 0.96% RRT 0.20% 0.96 1.09 0.96 RRT 0.04%RRT 0.18% RRT 0.90% 1.08 1.49 1.09 RRT 0.80% RRT 0.02% RRT 0.18% 1.091.50 1.49 RRT 0.19% RRT 0.88% RRT 0.02% 1.49 2.05 1.50 RRT 0.01% RRT2.49% 1.50 2.07 Total  2.11%  5.25%  6.16% Non- BBI- 4000 by HPLC (%)

All formulations showed good stability, however fewer non-BBI-4000materials were identified in formulations where antioxidants propylgallate or BHT were absent from the formulation.

Further stability testing has been completed for a 3-month time-frame,using Formulation No. 84-1, tested at three temperatures: accelerated(40° C.), room temperature (25° C.), and refrigerated (about 4° C.).Formulation No. 84-1 was specifically prepared using the followingpreparation instructions:

-   a) Combine the hexylene glycol and ethanol in a suitable container    and mix.-   b) Add the citric acid and stir to dissolve.-   c) Add the active (BBI-4000) and stir to dissolve.-   d) Add the KLUCEL™ MF and stir to dissolve, to increase viscosity of    the product.-   e) Lastly, add the DIMETHICONOL BLEND 20 and briefly disperse.-   f) Homogenize the mixture of steps a) through e). For small batches,    homogenation can be carried out by passing/mixing between 2 syringes    connected with a micro-emulsifying needle. For larger batches, an    overhead or inline homogenizer may be required.

The results of the 3-month stability study are provided in Table VI,below:

TABLE VI Stability of Formulation A 84-1 Day/ 7 D- 14 D- 30 D- 30 D- 90D- 30 D- 90 D- Temperature 0 40° C. 40° C. 40° C. 5° C. 5° C. 25° C. 25°C. Assay BBI-4000 (%) 9.81 10.32 10.21 10.25 9.32 10.50 10.26 10.63Total Non-BBI-4000 1.82  2.12  2.12  3.48 2.77  2.35  3.29  3.87 by HPLC(%)

The formulation 84-1, having the formulation shown in Table VII, showedgood stability and was tested in vivo.

TABLE VII A 84-1 Component % (w/w) BBI-4000 10 KLUCEL ™ MF 1.25(Hydoxypropyl Cellulose) Hexylene Glycol 10 DIMETHICONOL BLEND 20 2.5Citric Acid, Anhydrous 0.1 Ethanol (200 proof) 76.15 (Anhydrous Ethanol)

In the following clinical study, the A 84-1 formulation above wasmodified slightly. For 5% and 10% BBI-4000 gels, respectively, 0.001%anhydrous citric acid was used and the amount of ethanol adjustedaccordingly (81.25% for the 5% gel and 76.25% for the 10% gel).

EXAMPLE 4 Clinical Study

Study BBI-4000-CL-101: A Single-Center, Randomized, Double-Blind,Vehicle Controlled Study to Evaluate the Safety and the Effect on SweatProduction of Topically Applied BBI-4000 Gel in Subjects withHyperhidrosis

Study Design and Inclusion Criteria

Study BBI-4000-CL-101 was a Phase 1, randomized, double-blind,vehicle-controlled study of BBI-4000 gel conducted in 24 subjects withaxillary hyperhidrosis. The study was conducted at a single center inthe Dominican Republic. This study was not conducted under a US IND, butwas undertaken in full compliance with applicable regulations of theDominican Republic and with good clinical practice guidelines.

The objectives of this exploratory study were to evaluate the safety,local tolerability, and the effects on sweat production of topicallyapplied BBI-4000 gel. A preliminary assessment of systemic exposurebased on the pharmacokinetics of BBI-4000 was also conducted followingtopical application of the gel.

The drug product used in this study was an anhydrous semi-transparentgel with a composition including BBI-4000, hydroxypropylcellulose,hexylene glycol, DIMETHICONOL BLEND 20, citric acid, and ethanol.

The study consisted of 2 consecutive cohorts, where Cohort 1 establishedacceptable tolerability of 5% BBI-4000 gel (applied to one axilla) priorto enrolling a separate group of subjects into Cohort 2:

-   Cohort 1: 6 subjects received 5% BBI-4000 gel in one axilla and    vehicle in the other once daily (at night) for 14 consecutive days,    based on a randomized, split-body design.-   Cohort 2: 18 subjects (6 in each treatment group) were randomized to    receive 5% BBI-4000 gel, 10% BBI-4000 gel, or vehicle (control) to    both axillae once daily (at night) for 14 consecutive days, based on    a parallel-group design.

Subjects were 18 to 45 years of age, in good general health, with adiagnosis of primary axillary hyperhidrosis based on the followingcriteria at baseline:

-   HDSS of 3 or 4 (HDSS=Hyperhidrosis Disease Severity Score)-   Gravimetric test indicating at least an average of 100 mg of sweat    production in each axilla in 5 minutes at rest (at 25° C. to 27° C.)-   Bilateral and symmetrical hyperhidrosis

Subjects with prior axillary use of botulinum toxin (within 2 years) orreceiving any anticholinergic medication were not eligible toparticipate in the study. All female subjects of child-bearing potentialwere required to use a medically acceptable method of contraceptionwhile on active treatment.

Subjects were not allowed to use any antiperspirant 7 days prior tobaseline assessments and for the duration of the study.

Study Assessments and Endpoints Assessment of Local Tolerability

Local tolerability to topical BBI-4000 was assessed by the investigator(erythema, dryness, and scaling) and study subjects (burning anditching).

The investigator graded the severity of erythema, dryness, and scalingfor each axilla based on a 4-point scale, where “0” was absent, “1” wasminimal (barely perceptible), “2” was mild, “3” was moderate, and “4”was severe.

Subjects graded the severity of any burning or itching based on a4-point scale, where “0” was absent, “1” was minimal (an awareness, butno discomfort), “2” was mild, “3” was moderate, and “4” was severe.

-   Assessment of Safety

Safety was assessed by AEs, serious AEs (SAEs), or unexpected AEs; vitalsigns (blood pressure and heart rate); and clinical laboratory measures(hematology, chemistry, and urinalysis). Clinically relevant laboratoryfindings were to be collected as AEs (AEs=adverse events).

-   Assessment of Efficacy

Efficacy was assessed by the change in gravimetrically measured sweatproduction and the change in hyperhidrosis disease severity scale (HDSS)from baseline to Day 15 (end of therapy).

For the gravimetric assessment, sweat production was measured by placingfilter paper (pre-weighed) on the axilla for 5 minutes while the subjectwas in a semi-reclining position at room temperature. The filter paperwas covered with plastic during exposure to the axilla, and was thenweighed following the 5-minute exposure period to calculate the amountof sweat produced.

For the HDSS, subjects rated the severity of their hyperhidrosis on a4-point scale (1, 2, 3, or 4) based on the level of interference withtheir daily activities. A score of 1 indicated that “my sweating isnever noticeable and never interferes with my daily activities” and ascore of 4 indicated that “my sweating is intolerable and alwaysinterferes with my daily activities”.

Key Results of Study BBI-4000-CL-101

All subjects completed the study, including the follow-up visit at Day16, and received 14 days of study treatment in accordance with the studyprotocol. All subjects were included in the analysis of studyassessments (evaluable population). The subjects in Cohort 1(split-body) had no AEs reported and tolerated well the 5% BBI-4000 geland vehicle with only minimal to mild dryness and erythema reported in acouple of subjects during the study.

The results from Cohort 2 subjects, who received study drug in bothaxillary areas in a parallel design, are considered the most informativedata from this study and are the focus of the following sections.

This was an exploratory study not powered to achieve statisticallysignificant differences in the efficacy parameters measured, but toprovide an early indication of safety, tolerability, and the potentialtreatment effect of topically applied BBI-4000.

Baseline Demographics and Disease Characteristics

Subjects in Cohort 2 ranged from 18.6 to 43.7 years of age, with amedian age of ≤31 years in each treatment group. All subjects wereHispanic/Latino. No imbalances were noted between treatment groups withregard to gender, race, or ethnicity.

Measures of sweat production at baseline were generally similar betweentreatment groups and consistent with axillary hyperhidrosis. Mediansweat production was >200 mg (both axillae) in a 5-minute period for alltreatment groups based on baseline gravimetric assessment. All subjectshad an HDSS score of 3 or 4 at baseline.

Local Tolerability

Investigator and subject-based assessments of local tolerabilityindicated that 5% and 10% BBI-4000 gel topically applied to the axillaregion was well tolerated over the 14-day treatment period. Dryness,erythema, itching and burning were occassionally reported by 1 or 2subjects, they were minimal and did not lead to discontinuation of thetherapy in any individual.

Safety

No AEs were reported by any subjects during the conduct of the study,and no deaths or serious AEs were reported.

There were no changes in laboratory parameters that were consideredclinically relevant through the follow-up period (Day 16), as indicatedby no reports of laboratory-related AEs by the investigator.

There were no clinically relevant changes from baseline in vital signs(blood pressure and heart rate) for any treatment group in either cohortduring the study.

Efficacy

BBI-4000 formulation showed achievement of a greater reduction ingravimetrically measured sweat production and a greater improvement inHDSS assessments, when compared to vehicle. Although the overallreduction in sweat production and the HDSS improvement endpoints suggestthan BBI-4000 10% gel performed better than BBI-4000 5% gel, it isdifficult to make a definitive conclusion regarding differences in themagnitude of effect of the 2 active arms with this small sample size.Results for the key endpoints that have been commonly associated with aclinically meaningful improvement (i.e., reduction in sweat product ofat least 50% and ≥2-point improvement in HDSS) are here provided for theaggregate number of subjects exposed to BBI-4000 in comparison tovehicle.

The proportion of subjects treated with BBI-4000 who had at least a 50%reduction in sweat production at Day 15 was 75% (9 of 12) compared with33% (2 of 6) of subjects who received vehicle. In addition 8 of 12 (67%)subjects receiving BBI-4000 reported a 2-point improvement in HDSS atDay 15, compared with 2 of 6 (33%) in the vehicle group. This reductionin HDSS score represents a meaningful change from intolerable or barelytolerable hyperhidrosis to tolerable or never noticeable hyperhidrosisfor these subjects.

EXAMPLE 5 Further Anhydrous Formulations

Other formulations were developed, as set forth in the following table.

TABLE VIII Composition (% w/w) Component Gel 2 Gel 3 Gel 4 Gel 5 Gel 6Hydropropyl 1.250 1.250 1.250 1.250  1.250 Cellulose BBI-4000 5.0005.000 5.000 5.000  5.000 Hexylene 10.000 10.000 10.000 10.000 10.000Glycol Citric Acid, 0.001 0.001 0.001 0.001  0.001 Anhydrous Talc 0.500— — — — C12-15 — 2.500 — — — Alkyl Lactate Dimethicone — — 1.000 — —copolyol Magnesium — — — 0.001 — stearate Isopropyl — — — —  2.500myristate (IPM) Ethanol, qs to 100 qs to 100 qs to 100 qs to 100 qs to100 Anhydrous Visual Translucent, Clear Clear Clear Clear appearancemed-high colorless, colorless, colorless, colorless, at t = 0 viscosity,low low low low smooth viscosity, viscosity, viscosity, viscosity,smooth smooth smooth smooth Assessed for √ √ √ √ √ short-term stability

By way of example, the amount of BBI-4000 can alternatively be 10.000,15.000 or 20.000 (% w/w), with the quantity of anhydrous ethanoladjusted accordingly.

Other formulations in accord herewith are listed in Table IX below.

TABLE IX Composition (% w/w) Component Gel AA Get BB Gel CC Gel DD GelEE Hydropropyl 1.250 1.250  1.250  1.250  1.250 Cellulose BBI-400010.000 10.000 10.000 10.000 10.000 Hexylene 10.000 10.000 10.000 10.00010.000 Glycol Citric Acid, 0.001 0.001  0.001  0.001  0.001 AnhydrousDimethicone, 1.000 0.500 —  1.000  1.000 e.g. 350 cSt Cyclomethicone,1.000 1.000  1.000 — — e.g. 5-NF Isopropyl — —  2.500 —  2.500 myristateMyristyl — — —  2.500 — propionate Ethanol, qs to 100 qs to 100 qs qs qsAnhydrous to 100 to 100 to 100By way of example, the amount of BBI-4000 can alternatively be 5.000%w/w, 15.000% w/w or 20.000% w/w, with the amount of anhydrous ethanoladjusted accordingly.

The following is a general formulation for particularly preferredembodiments of the subject composition.

Formulation

Component Amount BBI-4000 1% to 20% w/w Hexylene Glycol   10% w/wHydroxypropyl Cellulose  1.25% w/w Citric Acid, Anhydrous 0.001% w/wDimethiconol Blend 20 or  2.5% w/w Isopropyl Myristate Anhydrous Ethanolqs to 100% w/wAdditional preferred embodiments include dimethicone or cyclomethicone,separately or in combination, in place of Dimethiconol Blend 20,optionally together with isopropyl myristate, in the above formulation.

In these compositions, isopropyl myristate is preferred overDimethiconol Blend 20 for two reasons. The first reason is related totransfer and scale-up of manufacturing process. It has been founddifficult to stabilize the droplets of Dimethiconol Blend 20 in theformulation when attempting to increase the batch size and transfer themanufacturing process. Over time, a small amount of Dimethiconol Blend20 coalesces at the bottom of the container in small droplets. Thechange to isopropyl myristate (IPM) eliminates this. The second reasonis related to FDA acceptance of Dimethiconol Blend 20. WhileDimethiconol Blend 20 is an acceptable ingredient in cosmeticpreparations, it has not been previously approved in pharmaceuticalformulations. The FDA accepted its use in clinical studies, butadditional studies might be needed to qualify it in a final formulation.The change to isopropyl myristate eliminates the need to conduct thisadditional testing as IPM has been approved in other pharmaceuticalpreparations, Dimethiconol Blend 20 was not detrimental to the functionof the formulation; however, isopropyl myristate provides benefits toscale-up and commercialization. After evaluating a number ofalternatives, isopropyl myristate was selected based upon its ability toreduce tack during drying as well as provide similar in vitropermeability. It also provided a formulation with a similar chemicalstability profile to Dimethiconol Blend 20. The isopropyl myristateformulation was compared to the Dimethiconol Blend 20 formulation in apreclinical animal study. The isopropyl myristate formulationdemonstrated an increase in permeation in mini-pigs relative to theDimethiconol Blend 20 formulation. Two upcoming studies in humans areplanned. The first study will compare the pharmacokinetics of 5% and 15%BBI-4000 gels containing isopropyl myristate with 15% BBI-4000 gelcontaining Dimethiconol Blend 20. The second study will evaluate/confirmthe efficacy of BBI-4000 gel containing isopropyl myristate.

While certain preferred and alternative embodiments have been set forthfor purposes of disclosure, modifications to the disclosed embodimentsmay occur to those who are skilled in the art. Accordingly, thisspecification is intended to cover all embodiments and combinations andmodifications thereof which do not depart from the spirit and scope ofthe following claims.

1.-23. (canceled)
 24. An anhydrous topical gel composition comprising:(a) a compound having the formula:

said compound having the R stereoisomeric configuration at the 2position and the R, S or RS stereoisomeric configuration at the 1′ and3′ positions, or being a mixture thereof; (b) anhydrous ethanol; (c) atleast one gelling or viscosity-controlling ingredient comprisinghydroxypropyl cellulose; (d) citric acid; (e) hexylene glycol; (f)isopropyl myristate; and (g) optionally, at least one additional carrieror excipient provided that said anhydrous topical gel composition isanhydrous and comprises from about 1% to about 25% w/v or w/w of thecompound of formula (2), said anhydrous topical gel composition havinggreater storage stability compared to a composition comprising anaqueous solvent or aqueous buffer, with the proviso that saidingredients are present in amounts such that the product of anytransesterification is the same as the compound of formula (2), and/orwith the proviso that said anhydrous ethanol is present in an amountsufficient to act as a non-aqueous solvent for the compound of formula(2).
 25. The anhydrous topical gel composition of claim 24, wherein theanhydrous topical gel composition comprises at least about 70% w/wanhydrous ethanol.
 26. The anhydrous topical gel composition of claim25, wherein the anhydrous topical gel composition comprises from about70% to about 99.99% w/w anhydrous ethanol.
 27. The anhydrous topical gelcomposition of claim 26, wherein the anhydrous topical gel compositioncomprises from about 70% to about 85% w/w anydrous ethanol.
 28. Theanhydrous topical gel composition of claim 24, wherein the anhydroustopical gel composition comprises at least one additional carrier orexcipient.
 29. The anhydrous topical gel composition of claim 24,wherein the compound of formula (2) is selected from the groupconsisting of: (i)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (ii) (2R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (iii) (2R,3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (iv) (2R,3′S)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (v) (2R,1′R,3′S)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (vi) (2R,1′S,3′S)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (vii) (2R,1′R,3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; and (viii) (2R,1′S,3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide.
 30. The anhydrous topical gel composition of claim 24, whereinthe anhydrous topical gel composition comprises the compound of formula(2) at a concentration of from about 1% w/v or w/w to about 20% w/v orw/w.
 31. The anhydrous topical gel composition of claim 30, wherein theanhydrous topical gel composition comprises the compound of formula (2)at a concentration of from about 2% w/v or w/w to about 10% w/v or w/w.32. The anhydrous topical gel composition of claim 31, wherein theanhydrous topical gel composition is packaged in a multiple dosecontainer that meters a dose of from about 0.5 ml to about 1.0 ml of thecomposition for each application.
 33. The anhydrous topical gelcomposition of claim 24, wherein the anhydrous topical gel compositionis packaged in a single or unit dose container that delivers a single orunit dose of about 0.5 ml to about 1.0 ml of the composition for eachapplication.
 34. The anhydrous topical gel composition of claim 24,wherein the anhydrous topical gel composition further comprises at leastone member selected from the group consisting of dimethicone andcyclomethicone.
 35. The anhydrous topical gel composition of claim 24,wherein the anhydrous topical gel composition further comprises a 6%silicone gum blend in dimethicone.
 36. The anhydrous topical gelcomposition of claim 24, wherein said anhydrous topical gel compositionis topically administered to an affected skin area of the subject,within 1-2 hours prior to the subject's sleep period.
 37. The anhydroustopical gel composition of claim 24, wherein said anhydrous topical gelcomposition is topically administered in a one to four times dailyregimen to an affected skin area of a subject.
 38. A method of treatinghyperhidrosis in a subject, said method comprising topicallyadministering an anhydrous topical gel composition to skin of an area ofa subject suffering from hyperhidrosis, before bedtime, such that,compared to untreated, baseline conditions, sweat production is reducedby at least 25% for at least six (6) hours; and such that sweatproduction is reduced by an amount substantially equivalent to an amountthat sweat production is reduced as compared to untreated, baselineconditions, following administration of a composition comprising thesame concentration of glycopyrrolate, and with an improved safetyprofile compared to topical glycopyrrolate, wherein said anhydroustopical gel composition comprises the following ingredients: (a) acompound having the formula:

said compound having the R stereoisomeric configuration at the 2position and the R, S or RS stereoisomeric configuration at the 1′ and3′ positions, or being a mixture thereof; (b) anhydrous ethanol; (c) atleast one gelling or viscosity-controlling ingredient comprisinghydroxypropyl cellulose; and (d) citric acid; (e) hexylene glycol; (f)isopropyl myristate; and (g) optionally, at least one additional carrieror excipient provided that said anhydrous topical gel composition isanhydrous and comprises from about 1% to about 25% w/v or w/w of thecompound of formula (2), said anhydrous topical gel composition havinggreater storage stability compared to a composition comprising anaqueous solvent or aqueous buffer, with the proviso that saidingredients are present in amounts such that the product of anytransesterification is the same as the compound of formula (2), and/orwith the proviso that said anhydrous ethanol is present in an amountsufficient to act as a non-aqueous solvent for the compound of formula(2), wherein the anhydrous topical gel composition is administered viadermal transmission by application to an anatomic area selected from ahand palm area, a foot plantar area, a groin area, an axilla area, and afacial area of the subject.
 39. The method of claim 38, wherein theanhydrous topical gel composition comprises at least about 70% w/wanhydrous ethanol.
 40. The method of claim 39, wherein the anhydroustopical gel composition comprises from about 70% to about 99.99% w/wanhydrous ethanol.
 41. The method of claim 40, wherein the anhydroustopical gel composition comprises from about 70% to about 85% w/wanydrous ethanol.
 42. The method of claim 38, wherein the anhydroustopical gel composition comprises at least one additional carrier orexcipient.
 43. The method of claim 38, wherein the compound of formula(2) is selected from the group consisting of: (i)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (ii) (2R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (iii) (2R,3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (iv) (2R,3′S)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (v) (2R,1′R,3′S)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (vi) (2R,1′S, 3′S)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; (vii) (2R,1′R,3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide; and (viii) (2R,1′S,3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide.
 44. The method of claim 38, wherein the anhydrous topical gelcomposition comprises the compound of formula (2) at a concentration offrom about 1% w/v or w/w to about 20% w/v or w/w.
 45. The method ofclaim 44, wherein the anhydrous topical gel composition comprises thecompound of formula (2) at a concentration of from about 2% w/v or w/wto about 10% w/v or w/w.
 46. The method of claim 38, wherein theanhydrous topical gel composition is packaged in a multiple dosecontainer that meters a dose of from about 0.5 ml to about 1.0 ml of thecomposition for each application.
 47. The method of claim 38, whereinthe anhydrous topical gel composition is packaged in a single or unitdose container that delivers a single or unit dose of about 0.5 ml toabout 1.0 ml of the composition for each application.
 48. The method ofclaim 38, wherein the compound of formula (2) is (2R, 3′R)3-(2-cyclopentyl-2-phenyl-2-hydroxyacetoxy)-1-(ethoxycarbonylmethyl)-1-methylpyrrolidiniumbromide.
 49. The method of claim 38, wherein the anhydrous topical gelcomposition further comprises at least one member selected from thegroup consisting of dimethicone and cyclomethicone.
 50. The method ofclaim 38, wherein the anhydrous topical gel composition furthercomprises a 6% silicone gum blend in dimethicone.
 51. The method ofclaim 38, wherein said anhydrous topical gel composition is topicallyadministered to an affected skin area of the subject, within 1-2 hoursprior to the subject's sleep period.
 52. The method of claim 38, whereinsaid anhydrous topical gel composition is topically administered in aone to four times daily regimen to an affected skin area of a subject.